Effects of Seed Priming on Vitality and Preservation of Pepper Seeds
Round 1
Reviewer 1 Report
The publication proposal is clear and does not raise any doubts. The first part introduces the reader fairly well, but only a few recently published works are included in the references. Methodically, the study was well prepared and described. Nevertheless, it would be interesting to determine the optimal concentration of KNO3 and PEG for seed treatment. There is no information, why only 3% KNO3, 20% PEG and 3% KNO3 + 20% PEG were used? Additionally, there will be interesting to get information about others priming solutions, not only the most commonly used.
The authors point to the differences in the obtained results between the 3 lines of peppers (No.63, No.73 and No.101), but there are lack of further information about used material (passport data or genebank accession no.). Why only these three lines were chosen?
These studies are well prepared, but should be consider as a preliminary research which should be extended. There are no conclusions regarding the observed differences, both in terms of genetics and biochemistry (structure of the seed coat).
Author Response
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Author Response File: 
Author Response.docx
Reviewer 2 Report
In this manuscript, the authors investigated the effects of different priming treatments (3% KNO3, 20% PEG, or 15 3% KNO3 + 20% PEG) vs- non-priming and 8 subsequent months storage on three pepper strains (No.63, No.73, and No.101).
The subject of research is interesting and the abstract effective, except for:
Line 17-18: it must be clarified that the germination percentage did not vary. Therefore, “Notwithstanding the germination percentage of primed seeds did not vary compared to non-primed ones, seeds germinated quickly and had strengthened emergence, especially for those primed….”
The introduction is presented in context with relevant literature and the aim of the paper both in the abstract and at the end of the introduction is well defined. However, from the premises of the introduction, I would have expected authors to make preliminary tests to choose the best concentration of KNO3 to use. In fact, the authors stated that the best concentration found for pepper seeds was 0.2-0.3% of KNO3 while 3% was the best one for seeds of other plant species. Moreover, they also discussed the high costs of PEG priming. Since the production of KNO3 is expensive and is further increasing due to the increase in the price of oil, why was not previously verified whether an intermediate concentration between 0.3 and 3% could still be effective? The authors should discuss this point.
The authors used a one-way ANOVA to analyze the effect of the priming treatments on the germinability for different pepper varieties. However, even non-taking into account the 0, 2, 4, 6, and 8 months storage, they should have used at least a two-way analysis of variance to consider strains x treatments, and a three-way ANOVA when to consider strains x treatments x types of bag/temperature of conservation. They can certainly use the same SPSS software version 21, applying , after the two or three-way ANOVA, the Tukey multiple comparison test for the mean separations at 0.05 probability level and show in the tables the significance of the interactions. After applying the correct statistical analysis, the results and discussions will certainly change and must be corrected.
Moreover, the MS would certainly improve if a principal component analysis (PCA) would be applied to the several parameters analyzed to identify a correlation between variables and visualize the real differences among strains, priming treatments, and types of bag/temperature.
Author Response
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Reviewer 3 Report
In the manuscript entitled: "Effects of seed priming on vitality and preservation of pepper seeds" (Number: agriculture-1661125), Authors present research on pepper seeds treatment with various substances in order to improve the germinability and viability of the seedlings of this plant. It is worth noting that the authors analyzed the storage parameters of primed seeds which is important for plants producers, especially those grown on an industrial scale. In my opinion, this topic is important from the economic point of view, especially in the context of the disappearance of the treatment effects during storage and the observed shortened life of the treated seeds in some plants species or cultivars.
I found some of problematic points in the manuscript, I only cite some of them below.
- In the Materials and Methods chapter there is no description of the seed breeding parameters used during the experiment.
- line # 124: “Unaged seed served as the control.”– please explain what "unaged" means.
- line # 155: “…and the seedlings appeared more robust.” - please explain what "robust" means.
- line # 240: “…the accelerated aging test was carried out.” – a more detailed description of this test should be provided.
In conclusion, in my opinion, used techniques are simple but acceptable. The scientific value of the research presented in the manuscript is very low, but the topic is interested from the economic point of view.
Author Response
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Round 2
Reviewer 1 Report
I find these results interesting but should be considered as preliminary research and should be extended to publication. The article is correct, therefore acceptable, but the content as it stands is of little significance.
Author Response
We thank the reviewer for your time and patience in helping us to improve our manuscript. We will make more efforts on priming treatments in future studies.
Reviewer 2 Report
The authors addressed all my concerns and, even if they did not perform a principal component analysis they explained the reason for not doing it. I am satisfied with the corrections.
Author Response
We gratefully thank the reviewer for the precious time spent making constructive remarks helping us to improve our manuscript.
This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.
Round 1
Reviewer 1 Report
Materials and Methods:
The experiment is expected to be repeated to obtain more reliable information (considering variation in results etc.).
Discussion:
- "the effect on the final germination percentage of seeds was not obvious" (line 230)- However, the opposite case may be for some crops. Amjad et. al. (2007) refer to this for pepper as; "Final germination percentage (FGP) of pepper seeds was significantly improved by different priming treatments over the control." Please reconsider the issue.
- There is a significant difference between pepper seeds lots. However, this issue was not mentioned enough in the result section and the reasons for the difference are not discussed.
- As mentioned in the conclusions section: "Different storage conditions had variable effects on primed pepper seed vigor (line 282)." However, the reasons for this situation have not been sufficiently emphasized and have not been discussed. The reasons for the difference among the seed lots (for storage conditions) have not been sufficiently mentioned as well.
Author Response
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Reviewer 2 Report
The manuscript entitled “Effects of seed priming on vitality and reservation of pepper seeds” analyzed the effect of three methodologies of priming on pepper seeds used for 7 days or 14 days to verify the homogeneity of germination and the storage (until 8 months) of seeds after these priming methodologies and different methodologies and environment of conservation (plastic bag at 25 °C; plastic bag at -4°C, paper bag at 25°C).
I think that the experiment does not enhance the knowledge about the topic because this study resulted a replication of experiments already conducted by other authors. Moreover, I have some concerns about the manuscript:
Abstract
Lines 15: “a series of experiments”. Which experiments? I think that authors must specify the priming treatments, otherwise the abstract cannot support itself, but readers must read the article.
Line 18: specify the length of storage.
Keywords
Eliminate the numbers.
Introduction
Line 31: I suggest to introduce a reference to confirm this sentence.
Lines 40-41: Why is the PG difficult to infiltrate into seeds?
Lines 51-52: I don’t understand why authors have written that the time of osmotic adjustment was 12-24 h. Indeed, this affirmation is not necessary to the aim of the study and the time of osmotic adjustment after KNO3 treatment was not verified by these authors.
Line 63: eliminate the comma between reduced and compared.
Lines 67-68: “higher temperature and humidity conditions” compared to what? Which range of temperature and relative humidity?
Line 69: what is the meaning of “mild” in this sentence? Who decided if the conditions are mild? Authors, references cited before of this sentence used similar temperature of the authors of reference 23.
Lines 73-76: I don’t understand which information this sentence want to do.
Materials and Methods
Line 104: -4°C or +4°C? Have authors froze the seeds? Was a refrigerator or a freezer? Specify the brand and the city of the refrigerator.
Line 107-108: specify the length of storage.
Line 109: what’s the meaning of this part of experiment? It was not introduced before in the introduction section or in the aim by authors
Line 110: the meaning of the word “dish” is food recipe; the word to refer to an instrument is “plate”.
Line 114: which kind of ANOVA was done? One, two or three anova? Which variability factors were used?
Results
I cannot understand the differences between treatments and methodologies of storage through Table and Figures present in this manuscript because they are devoid of a statistical analysis? What authors were based on to write results?
Figures 3-4: trough this figures the difference between priming effect at 7th day and at 14th day is not clear. I suggest a summarize of these results to underline these differences.
Figure 4: it is not evident the comparison between the effect of different storage treatments and the different priming methodologies.
Discussion
Lines 235-236: this is an opinion of authors or they were based on viscosity values?
Line 238: PEG-6000 was not present before throughout the manuscript.
Line 242: Howe can the cost be reduced if we must buy two primers instead of one, and the only KNO3 led to better results than the combination of the two primers?
Line 249: the reference Saddam et al. is non present in the reference list.
A real discussion is missing and the mainly references on this topic are missing in this section.
Author Response
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Author Response File: Author Response.docx
